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抗铜绿假单胞菌血清型菌株单克隆抗体的制备与鉴定

Production and characterization of monoclonal antibodies against serotype strains of Pseudomonas aeruginosa.

作者信息

Lam J S, MacDonald L A, Lam M Y, Duchesne L G, Southam G G

出版信息

Infect Immun. 1987 May;55(5):1051-7. doi: 10.1128/iai.55.5.1051-1057.1987.

Abstract

Monoclonal antibodies against 12 of the 17 IATS serotype strains of Pseudomonas aeruginosa were produced. Eighty-seven hybridoma clones were isolated, and the antibodies secreted were found to be reactive with both Formalin-fixed whole cells and purified lipopolysaccharide of homologous strains in enzyme-linked immunosorbent assays. Among these monoclonal antibodies, the predominant antibody class was immunoglobulin M (IgM) (76%), although antibodies of the IgG2a and IgG3 isotypes were also produced. The monoclonal antibodies could further be divided into two groups based on their ability to agglutinate whole cells of homologous strains. The agglutinating monoclonal antibodies were found to immunoblot with the O side chains of homologous lipopolysaccharide, while the nonagglutinating monoclonal antibodies were found to be reactive with outer membrane protein-associated lipopolysaccharide. The applicability of monoclonal antibodies for serotyping was examined, and several antibodies were found to agglutinate whole cells and immunoblot with the O antigen of corresponding serotypes of clinical isolates from cystic fibrosis patients. In conclusion, a set of monoclonal antibodies against the IATS serotype strains of P. aeruginosa have been produced. These antibodies represent a bank of invaluable immunological reagents which may have application in serotyping, epitope mapping, lipopolysaccharide structural determination, and studies of protection against P. aeruginosa.

摘要

制备了针对17株铜绿假单胞菌国际抗血清分型(IATS)血清型菌株中12株的单克隆抗体。分离出87个杂交瘤克隆,在酶联免疫吸附试验中发现所分泌的抗体与福尔马林固定的全细胞以及同源菌株的纯化脂多糖均有反应。在这些单克隆抗体中,主要的抗体类别是免疫球蛋白M(IgM)(76%),不过也产生了IgG2a和IgG3同种型的抗体。根据单克隆抗体凝集同源菌株全细胞的能力,可将其进一步分为两组。发现具有凝集作用的单克隆抗体可与同源脂多糖的O侧链进行免疫印迹分析,而不具有凝集作用的单克隆抗体则与外膜蛋白相关脂多糖有反应。检测了单克隆抗体在血清分型中的适用性,发现几种抗体可凝集全细胞,并与来自囊性纤维化患者临床分离株相应血清型的O抗原进行免疫印迹分析。总之,已制备出一组针对铜绿假单胞菌IATS血清型菌株的单克隆抗体。这些抗体是一批非常宝贵的免疫试剂,可应用于血清分型、表位作图、脂多糖结构测定以及抗铜绿假单胞菌保护性研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a517/260467/3f7e266a0cfd/iai00089-0044-a.jpg

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